Archives of Pharmacal Research

  • 제21권5호
  • /
  • Pages.521-526
  • /
  • 1998
  • /
  • 0253-6269(pISSN)
  • /
  • 1976-3786(eISSN)
대한약학회 (The Pharmaceutical Society of Korea)
권호 표지

Physicochemical Properties of Recominant Hepatitis B Surface Antigen Expressed in Mammalian Cell(C127)

  • 발행 : 1998.10.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The physicochmical properties of recombinant hepatitis B surface antigen (r-HBsAg), which was expressed in C127 mammalian cell were studied. Using roller bottle culture in DMEM supplemented with fetal bovine serum, 10-15 mg/L of r-HBsAg was produced with about 31% of purification yield. The purity of r-HBsAg by HPLC was 99.8% and electron microscopic examination showed homogeneous spherical particle with 22 nm in diameter, a morphological characteristic of HBsAg. The density of r-HBsAg by CsCI density gradient method was 1.19g/ml and the isoelectric point by Mono $P^{TM}$ HR 5/20 column was 4.6. The analysis of subunit protein pattern using SDS-PAGE followed by scanning densitometry gave 81.3% of S protein and 18.7% of pre-S protein. fluorophore-assisted-carbohydrate-electrophoresis analysis showed the relative amount of carbohydrate to protein was 1.7% and it smajr component was N-acetyl glucosamine, which was about 39% of total carbohydrate. The relative amount of lipid to protein determined by vanillin phosphoric acid method was 32.5% and its major component was phospholipid, which was about 70% of total lipid. The physicochemical properties of C127 mammalian cell-derved r-HBsAg are similar to those of p-HBsAg, suggesting that the r-HBsAg can be used in developing a new preventive vaccine against hepatitis B.

키워드

참고문헌

  1. International Symposium on Viral Hepatitis and Liver Disease Clinical trial of a pre-S2 ontaining hepatitis B vaccine:Early response of anti pre-S2 in vaccins and efficacy in non-responders to conventional vaccins Akahane;Y.
  2. J.Chromatogr. v.175 High performance liquid chmatography of cholesterol autooxidation products Ansari;G.A.S.;Smith;L.
  3. viral hepatitis and liver disease Beasley;R.P.;Hwang;L.Y.
  4. Can. J. Biochem. Psysiol. v.37 A rapid method of total lipid extraction and purification Blogh;E.G.;Dyer;W.J.
  5. JAMA v.191 A new antigen in leukemia sera Blumberg;B.S.;Alter;H.J.;Visnich;S.
  6. Bull. N. Y. Acad. Med. v.44 Hepatitis and leukemia:their ralation to Australia antigen Blumberg;B.S.;Sutnick;A.L.;London;W.T.
  7. Vox Sang v.44 Distribution of HBsAg subtypes in the world Courouce-Pauty;A.M.;Plancon;A.;Soulier;J.P.
  8. Lancet v.1 Dane;D.S.;Camereron;C.H.;Briggs;M.
  9. Vaccine v.15 Comparison between hepatitis B surface antigen (HBsAg)particles derived from mammalian cells (CHO) and yeast cells (Hansenula polymorpha):compositon;structure and immunogenicity Diminsky;D.;Schirmbeck;R.;Reimann;J.;Barenholz;Y.
  10. Biochem. J. v.265 Hepatitis B syrface antigen. Role of lipids in maintaining the structutal and antigenic properties of protein components Gavilanes;F.;Gomez-Gutierrez;J.;Araccil;M.;Gonzalez-Ros;J.M.;Ferragut;J.A.;Guerrero;E.;Peterson;D.L.
  11. J. Hepatol v.17 no.SUP.3 Studies on the attachment and penetration of hepatitis B virus Gerlich;W.H.;Lu;X.;Heermann;K.H.
  12. J. Virol. v.52 Large surface proteins of hepatitis B virus containing the pre-S sequence Heerman;K.H.;Goldmann;U.;Schwartz;W.;Seyffarth;T.;Baumgarten;H.;Gerlich;W.H.
  13. J. Viral. Hepat. v.3 Reactogenicity and imunogenicity of a new recombinant hepatitis B vaccine containing Pre S antigens:a prelominary report Hourvitz;A.;Mosseri;R.;Solomon;A.;Yehezkelli;Y.Atsmin;J.Danon;Y.L.;Koren;R.;Shouval;D.
  14. Progress in hepatitis B immunization v.194 Significance of pre-antigens for hepatitis B vaccines Krone;B.;Gerlich;W.H.;Heerman;K.H.;Lenz;A.;Lu;S.;Seiter;M.;Tomslen;R.
  15. Biotechnology v.3 Expression of amplified gepatitis B virus surface antigen genes in chinese hamster ovary cells Michel;M.L.;Sobczak;E.;Malpiece;Y.;Tiollais;P.;Streek;R.E.
  16. Proc. Natl. Acad. Sci. USA. v.81 Synthesis in animal all of hepatitis B surface antigen particles carrying a receptor for polymerized human serum albumun Michel;M.L.;Pontisso;P.Sobzak;E.;Malpiece;Y.Streek;R.E.;Tiollais;P.
  17. Vaccine v.4 Antibodies to a synthetic peptide from the pre-S 120-145 region of the hepatitis B virus envelope are virus neutralizing Neurath;A.R.
  18. Vaccine v.7 Expression of hepatitis B virus surface antigen containing the pre-S region in mammalian cell culture system Samanta;H.;Youn;B.W.
  19. J. Hepatol. v.17 no.SUP.3 Fifth international symposium on viral hepatitis summary Shelock;S.
  20. J. Chromatogr. A. v.720 Fluoropfore-assisted carbohydrate electrophoresis in the separation;analysis;and sequencing of carbohydrates Starr;C.M.;Masada;R.I.;Hague;C.;Skop;E.;Klock;J.C.
  21. J. Virol. v.46 Stuctural relationships between minor and major proteins of hepatitis B surface antigen Stibbe;W.;Gerlich;W.H.
  22. Progress in hepatitis B immunization v.194 no.35 Evaluation of immune responses in a third generation hepatitis B vaccine containing pre-S proteins in comparative trials Thoma;H.A.;Hemmerling;A.E.;Hotzinger;H.
  23. Nature v.317 The hepatitis B virus Tiollais;P.Paucel;C.;Dejean;A.
  24. Nature v.298 Synthesis and assembly of hepatitis B virus surface antigen particles in yeast Valenzuela;P.;Medina;A.;Rutter;W.J.;Ammerer;G.;Hall;B.D.